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Molecular and Cellular Biology

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https://www.readbyqxmd.com/read/27895153/ubiquitylation-of-ku80-by-rnf126-promotes-completion-of-nhej-mediated-dna-repair
#1
Noriko Ishida, Tadashi Nakagawa, Shun-Ichiro Iemura, Akira Yasui, Hiroki Shima, Yasutake Katoh, Yuko Nagasawa, Toru Natsume, Kazuhiko Igarashi, Keiko Nakayama
Repair of damaged DNA is critical for maintenance of genetic information. In eukaryotes, DNA double-strand breaks (DSBs) are recognized by the Ku70-Ku80 heterodimer, which then recruits proteins that mediate repair by nonhomologous end-joining (NHEJ). Prolonged retention of Ku70/80 at DSBs prevents completion of repair, however, with ubiquitylation of Ku80 having been implicated in Ku70/80 dissociation from DNA. Here we identify RNF126 as a ubiquitin ligase that is recruited to DSBs and ubiquitylates Ku80, with UBE2D3 serving as an E2...
November 28, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27895152/spatio-temporal-uncoupling-of-mirna-mediated-translational-repression-and-target-rna-degradation-controls-mirnp-recycling-in-mammalian-cells
#2
Mainak Bose, Bahnisikha Barman, Avijit Goswami, Suvendra N Bhattacharyya
miRNA-mediated repression control expression of more than half of protein coding genes in metazoan animals. Translation repression is associated with target mRNA degradation initiated by decapping and deadenylation of the repressed mRNAs. Earlier evidence suggest Endoplasmic Reticulum (ER) as the site where miRNPs interact with their targets before the translation repression sets in but the subcellular location of subsequent degradation of miRNA-repressed messages is largely unidentified. Here, we explore the subcellular distribution of essential components of degradation machineries of miRNA-targeted mRNAs...
November 28, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27895151/high-fat-diet-promotes-mammary-gland-myofibroblast-differentiation-through-mir-140-downregulation
#3
Benjamin Wolfson, Yongshu Zhang, Ramkishore Gernapudi, Nadire Duru, Yuan Yao, Pang-Kuo Lo, Qun Zhou
Human breast adipose tissue is a heterogonous cell population consisting of mature white adipocytes, multipotent mesenchymal stem cells, committed progenitor cells, fibroblasts, endothelial cells, and immune cells. Dependent on external stimulation, adipose derived stem cells differentiate along diverse lineages into adipocytes, chondrocytes, osteoblasts, fibroblasts, and myofibroblasts. It is currently not fully understood how high-fat diet reprograms adipose-derived stem cells into myofibroblasts. In our study, we use regular-diet and high-fat diet induced obesity mouse models to investigate the role of dietary fat on myofibroblast differentiation in the mammary stromal microenvironment...
November 28, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27872149/endogenous-tert-n-terminal-tagging-affects-human-telomerase-function-at-telomeres-in-vivo
#4
Kunitoshi Chiba, Jacob M Vogan, Robert A Wu, Manraj S Gill, Xiaozhu Zhang, Kathleen Collins, Dirk Hockemeyer
Telomerase action at telomeres is essential for the immortal phenotype of stem cells and the aberrant proliferative potential of cancer cells. Insufficient telomere maintenance can cause stem cell and tissue failure syndromes, while increased telomerase levels are associated with tumorigenesis. Both pathologies can arise from only small perturbation of telomerase function. To analyze telomerase at its low endogenous expression level, we genetically engineered human pluripotent stem cells (hPSCs) to express various N-terminal fusion proteins of the telomerase reverse transcriptase from its endogenous locus...
November 21, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27872148/proteomics-screen-identifies-class-i-rab11-fips-as-key-regulators-of-cytokinesis
#5
Carl Laflamme, Jacob A Galan, Khaled Ben El Kadhi, Antoine Méant, Carlos Zeledon, Sébastien Carréno, Philippe P Roux, Gregory Emery
The 14-3-3 protein family orchestrates a complex network of molecular interactions that regulates various biological processes. Owing to their role in regulating the cell cycle and protein trafficking, 14-3-3 proteins are prevalent in human diseases, such as cancer, diabetes and neurodegeneration. 14-3-3 proteins are expressed in all eukaryotic cells, suggesting that they mediate their biological functions through evolutionarily conserved protein interactions. To identify these core 14-3-3 client proteins, we used an affinity-based proteomics approach to characterize and compare the human and Drosophila 14-3-3 interactome...
November 21, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27849571/arginine-methylation-by-prmt1-regulates-muscle-stem-cell-fate
#6
Roméo Sébastien Blanc, Gillian Vogel, Xing Li, Zhenbao Yu, Shawn Li, Stéphane Richard
Quiescent muscle stem cells (MSC) become activated in response to skeletal muscle injury to initiate regeneration. Activated MSCs proliferate and differentiate to repair damaged fibers or self-renew to maintain the pool and insure future regeneration. The balance between self-renewal, proliferation and differentiation is a tightly regulated process controlled by a genetic cascade involving determinant transcription factors such as Pax7, Myf5, MyoD, and MyoG. Recently there have been several reports about the role of arginine methylation as a requirement for epigenetic-mediated control of muscle regeneration...
November 14, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27849570/the-werner-syndrome-helicase-coordinates-sequential-strand-displacement-and-fen1-mediated-flap-cleavage-during-polymerase-%C3%AE-elongation
#7
Baomin Li, Sita Reddy, Lucio Comai
The Werner syndrome protein (WRN) suppresses the loss of telomeres replicated by lagging strand synthesis by a yet to be defined mechanism. Here we show that whereas either WRN or the Bloom syndrome helicase (BLM) stimulate DNA polymerase δ progression across telomeric G-rich repeats, only WRN promotes sequential strand displacement synthesis and FEN1 cleavage, a critical step in Okazaki fragment maturation, at these sequences. Helicase activity as well as the conserved winged-helix (WH) motif and the Helicase-and-RNaseD C-terminal (HRDC) domain play important but distinct roles in this process...
November 14, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27849569/suppressor-of-fused-chaperones-gli-proteins-to-generate-transcriptional-responses-to-sonic-hedgehog-signaling
#8
Ziyu Zhang, Longyan Shen, Kelvin Law, Zengdi Zhang, Xiaotong Liu, Hu Hua, Sanen Li, Huijie Huang, Shen Yue, Chi-Chung Hui, Steven Y Cheng
Cellular responses to the graded Sonic hedgehog (Shh) morphogenic signal are orchestrated by three Gli genes that give rise to both transcription activators and repressors. An essential downstream regulator of the pathway, encoded by the tumor suppressor gene Suppressor of fused (Sufu), plays critical roles in the production, trafficking and function of Gli proteins, but the mechanism remains controversial. Here, we show that Sufu is up-regulated in active Shh responding tissues and accompanies Gli activators translocating into and Gli repressors out of the nucleus...
November 14, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27821480/rbm4-regulates-neuronal-differentiation-of-mesenchymal-stem-cells-by-modulating-alternative-splicing-of-pyruvate-kinase-m
#9
Chun-Hao Su, Kuan-Yang Hung, Shih-Chieh Hung, Woan-Yuh Tarn
RBM4 promotes differentiation of neuronal progenitor cells and neurite outgrowth of cultured neurons via its role in splicing regulation. In this study, we further explored the role of RBM4 in neuronal differentiation. During neuronal differentiation, energy production shifts from glycolysis to oxidative phosphorylation. We found that the splice isoform change of the metabolic enzyme pyruvate kinase M (PKM) from PKM2 to PKM1 occurs during brain development and is impaired in RBM4-deficient brain. The PKM isoform change could be recapitulated in human mesenchymal stem cells (MSCs) during neuronal induction...
November 7, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27821479/functional-roles-of-acetylated-histone-marks-at-mouse-meiotic-recombination-hotspots
#10
Irina V Getun, Zhen Wu, Mohammad Fallahi, Souad Ouizem, Qin Liu, Weimin Li, Roberta Costi, William R Roush, John L Cleveland, Philippe R J Bois
Meiotic recombination initiates following the formation of DNA double strand breaks (DSBs) by the Spo11 endonuclease early in prophase I at discrete regions in the genome coined hotspots. In mammals, meiotic DSB site selection is directed in part by sequence specific binding of PRDM9, a polymorphic histone H3 (H3K4Me3) methyltransferase. However, other chromatin features needed for meiotic hotspot specification are largely unknown. Here, we show that the recombinogenic cores of active hotspots in mice harbor several histone H3 and H4 acetylation and methylation marks that are typical of open, active chromatin...
November 7, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27821478/mcl-1-depletion-impairs-dna-dsb-repair-and-re-initiation-of-stalled-dna-replication-forks
#11
Abid R Mattoo, Raj K Pandita, Sharmistha Chakraborty, Vijaya Charaka, Kalpana Mujoo, Clayton R Hunt, Tej K Pandita
Myeloid cell leukemia-1 : MCL-1) is a pro-survival BCL-2 protein family member highly expressed in hematopoietic stem cells (HSCs) and regulated by growth factor signals that manifest anti-apoptotic activity. Here we report that depletion of MCL-1, but not its isoform MCL1S, increases genomic instability and cell sensitivity to ionizing radiation (IR) induced death. MCL-1 association with genomic DNA increased post-irradiation and it co-localized with 53BP1 in foci. Post-irradiation, MCL-1 depleted cells exhibited decreased γ-H2AX foci, decreased phosphorylation of ATR and higher levels of residual 53BP1 and RIF1 foci, suggesting DNA DSB repair by homologous recombination (HR) was compromised...
November 7, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27821477/interchangeable-roles-for-e2f-transcriptional-repression-by-the-retinoblastoma-protein-and-p27kip1-cdk-regulation-in-cell-cycle-control-and-tumor-suppression
#12
Michael J Thwaites, Matthew J Cecchini, Daniel T Passos, Ian Welch, Frederick A Dick
The mammalian G1-S phase transition is controlled by the opposing forces of cyclin dependent kinases (CDK) and the retinoblastoma protein (pRB). Here we present evidence for systems level control of cell cycle arrest by pRB-E2F and p27-CDK regulation. By introducing a point mutant allele of pRB that is defective for E2F repression (Rb1(G)) into a p27(KIP1) null background (Cdkn1b(-/-)), both E2F transcriptional repression and CDK regulation are compromised. These double mutant Rb1(G/G); Cdkn1b(-/-) mice are viable and phenocopy Rb1(+/-) mice in developing pituitary adenocarcinomas, even though neither single mutant strain is cancer prone...
November 7, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27821476/notch-downregulation-and-extramedullary-erythrocytosis-in-hif-prolyl-4-hydroxylase-2-deficient-mice
#13
Mikko N M Myllymäki, Jenni Määttä, Elitsa Y Dimova, Valerio Izzi, Timo Väisänen, Johanna Myllyharju, Peppi Koivunen, Raisa Serpi
Erythrocytosis is mainly driven by erythropoietin, which is regulated by hypoxia-inducible factor (HIF). Mutations in HIF prolyl 4-hydroxylase 2 (HIF-P4H-2/PHD2/EGLN1), the major downregulator of HIFα subunits, are found in familiar erythrocytosis, and large-spectrum conditional inactivation of HIF-P4H-2 in mice leads to severe erythrocytosis. Although bone marrow is the primary site for erythropoiesis, spleen remains capable for extramedullary erythropoiesis. We studied HIF-P4H-2 deficient mice (Hif-p4h-2(gt/gt)) which show slightly induced erythropoiesis upon aging despite non-increased erythropoietin levels, and identified spleen as the site of extramedullary erythropoiesis...
November 7, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27821475/asc1p-rack1-connects-ribosomes-to-eukaryotic-phospho-signaling
#14
Kerstin Schmitt, Nadine Smolinski, Piotr Neumann, Samantha Schmaul, Verena Hofer-Pretz, Gerhard H Braus, Oliver Valerius
WD40-repeat proteins fold into characteristic β-propeller structures and control signaling circuits during cellular adaptation processes within eukaryotes. The RACK1 protein of Saccharomyces cerevisiae, Asc1p, exclusively consists of a single seven-bladed β-propeller that operates from the ribosomal base at the head region of the 40S subunit. Here, we show that the ribosome binding-compromised variant R38D K40E (Asc1DEp) gets severely destabilized through the mutation of phospho-site T143 to dephosphomimicking alanine, probably through proteasomal degradation, leading to asc1(-) phenotypes...
November 7, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27799292/sumo-modified-fadd-recruits-cytosolic-drp1-and-caspase-10-to-mitochondria-for-regulated-necrosis
#15
Seon-Guk Choi, Hyunjoo Kim, Eun Il Jeong, Sungwoo Park, Song-Yi Lee, Hyeon-Jeong Lee, Seong Won Lee, Chin Ha Chung, Yong-Keun Jung
Fas-associated protein with Death Domain (FADD) plays a key role in extrinsic apoptosis. Herein, we show that FADD is SUMOylated as an essential step during intrinsic necrosis. FADD was modified at multiple lysine residues (K120/125/149) by Small Ubiquitin-related Modifier 2 (SUMO2) during necrosis caused by calcium ionophore A23187 and by ischemic damage. SUMOylated FADD bound to dynamin-related protein 1 (Drp1) in cells both in vitro and in ischemic tissue damage cores, thus promoting Drp1 recruitment by Mitochondrial fission factor (Mff) to accomplish mitochondrial fragmentation...
October 31, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27777312/mbd4-facilitates-immunoglobulin-class-switch-recombination
#16
Fernando Grigera, Robert Wuerffel, Amy L Kenter
Immunoglobulin heavy chain class switch recombination (CSR) requires targeted formation of DNA double strand breaks (DSBs) in repetitive switch region elements followed by ligation between distal breaks. The introduction of DSBs is initiated by activation induced cytidine deaminase (AID) and requires base excision repair (BER) and mismatch repair (MMR). The BER enzyme, methyl CpG binding domain protein 4 (MBD4) has been linked to the MMR pathway through its interaction with MutL homologue 1 (MLH1). We find that when the Mbd4 exons 6-8 are deleted in a switching B cell line DSB formation is severely reduced and CSR frequency is impaired...
October 24, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27777311/glucocorticoid-receptor-accelerates-but-is-dispensable-for-adipogenesis
#17
Young-Kwon Park, Kai Ge
Dexamethasone (DEX), a synthetic ligand for glucocorticoid receptor (GR), is routinely used to stimulate adipogenesis in culture. GR-depleted preadipocytes show adipogenesis defects one week after induction of differentiation. However, it has remained unclear whether GR is required for adipogenesis in vivo By deleting GR in precursors of brown adipocytes, we found unexpectedly that GR is dispensable for brown adipose tissue development in mice. In culture, GR-deficient primary or immortalized white and brown preadipocytes showed severely delayed adipogenesis one week after induction of differentiation...
October 24, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27777310/distinct-roles-of-transcription-factors-klf4-krox20-and-ppar%C3%AE-in-adipogenesis
#18
Young-Kwon Park, Limin Wang, Anne Giampietro, Binbin Lai, Ji-Eun Lee, Kai Ge
Much of our knowledge on adipogenesis comes from cell culture models of preadipocyte differentiation. Adipogenesis is induced by treating confluent preadipocytes with the adipogenic cocktail, which activates transcription factors (TFs) glucocorticoid receptor (GR) and CREB within minutes and increases expression of TFs C/EBPβ, C/EBPδ, KLF4 and Krox20 within hours. All of these TFs have been shown to be capable of promoting adipogenesis in culture when they are overexpressed. However, it has remained unclear whether endogenous KLF4 and Krox20 are required for adipogenesis in culture and in vivo Using conditional knockout mice and derived white and brown preadipocytes, we show that endogenous KLF4 and Krox20 are dispensable for adipogenesis in culture and brown adipose tissue development in mice...
October 24, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27795300/eya1-s-conformation-specificity-in-dephosphorylating-phosphothreonine-in-myc-and-its-activity-on-myc-stabilization-in-breast-cancer
#19
Jun Li, Yoel Rodriguez, Chunming Cheng, Lei Zeng, Elaine Y M Wong, Chelsea Y Xu, Ming-Ming Zhou, Pin-Xian Xu
EYA1 is known to be overexpressed in human breast cancer in which Myc protein is also accumulated in association with decreased phospho-T58 levels. We have recently reported that EYA1 functions as a unique protein phosphatase to dephosphorylate Myc at phosphor-T58 to regulate Myc levels. However, it remains unclear whether EYA1-mediated Myc dephosphorylation on T58 1 is a critical function in regulating Myc protein stability in breast cancer. Furthermore, EYA1's substrate specificity has remained elusive. In this study, we have investigated these questions and here we report that depletion of EYA1 using shRNA in breast cancer cells destabilizes Myc protein and increases pT58 levels, leading to an increase in doubling time and impairment of cell cycle progression...
October 17, 2016: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/27795299/ifn%C3%AE-production-is-regulated-by-p38-mapk-in-macrophages-via-both-msk1-2-and-ttp-dependent-pathways
#20
Victoria A McGuire, Dalya Rosner, Olga Ananieva, Ewan A Ross, Suzanne E Elcombe, Shaista Naqvi, Mirjam M W van den Bosch, Claire E Monk, Tamara Ruiz-Zorrilla Diez, Andrew R Clark, J Simon C Arthur
Autocrine or paracrine signaling by the type I interferon IFNβ is essential for many of the responses of macrophages to pathogen-associated molecular patterns. This feedback loop contributes to pathological responses to infectious agents, and is therefore tightly regulated. We demonstrate here that macrophage expression of IFNβ is negatively regulated by mitogen- and stress-activated kinases (MSK) 1 and 2. Lipopolysaccharide- (LPS-) induced expression of IFNβ was elevated in both MSK1/2 knockout mice and macrophages...
October 17, 2016: Molecular and Cellular Biology
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